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Applied and Environmental Microbiology, August 2004, p. 4921-4929, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4921-4929.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

"Candidatus Endobugula glebosa," a Specific Bacterial Symbiont of the Marine Bryozoan Bugula simplex

Grace E. Lim and Margo G. Haygood*

Marine Biology Research Division and Center for Marine Biomedicine and Biotechnology, Scripps Institution of Oceanography, and Cancer Center, University of California, San Diego, La Jolla, California 92093-0202

Received 9 February 2004/ Accepted 5 May 2004

The bryozoans Bugula neritina and Bugula simplex harbor bacteria in the pallial sinuses of their larvae as seen by electron microscopy. In B. neritina, the bacterial symbiont has been characterized as a gamma-proteobacterium, "Candidatus Endobugula sertula." "Candidatus E. sertula" has been implicated as the source of the bryostatins, polyketides that provide chemical defense to the host and are also being tested for use in human cancer treatments. In this study, the bacterial symbiont in B. simplex larvae was identified by 16S rRNA-targeted PCR and sequencing as a gamma-proteobacterium closely related to and forming a monophyletic group with "Candidatus E. sertula." In a fluorescence in situ hybridization, a 16S ribosomal DNA probe specific to the B. simplex symbiont hybridized to long rod-shaped bacteria in the pallial sinus of a B. simplex larva. The taxonomic status "Candidatus Endobugula glebosa" is proposed for the B. simplex larval symbiont. Degenerate polyketide synthase (PKS) primers amplified a gene fragment from B. simplex that closely matched a PKS gene fragment from the bryostatin PKS cluster. PCR surveys show that the symbiont and this PKS gene fragment are consistently and uniquely associated with B. simplex. Bryostatin activity assays and chemical analyses of B. simplex extracts reveal the presence of compounds similar to bryostatins. Taken together, these findings demonstrate a symbiosis in B. simplex that is similar and evolutionarily related to that in B. neritina.

* Corresponding author. Mailing address: Scripps Institution of Oceanography, Marine Biology Research Division, 0202, University of California, San Diego, La Jolla, CA 92093-0202. Phone: (858) 534-5987. Fax: (858) 822-5839. E-mail: mhaygood{at}ucsd.edu.

Applied and Environmental Microbiology, August 2004, p. 4921-4929, Vol. 70, No. 8
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.8.4921-4929.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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